Anti-beta Tubulin抗体- Loading Control
参阅全部 beta Tubulin 一抗
兔多克隆抗体to beta Tubulin - Loading Control
Rabbit
This antibody detects a single clean band at 50kD representing beta Tubulin. This band is significantly reduced by using peptide blocking.
适用于: WB, ICC/IF, IHC-P, IPmore details
与反应: Mouse, Rat, Human, Chinese hamster
预测可用于: Chicken, Pig, Xenopus laevis, Zebrafish
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
(Peptide available as ab20775)
WB: HeLa, A431, MCF7, NIH3T3, PC12 CHO/K1, and 293 cell lysates; IP: HeLa whole cell extract; ICC/IF: HeLa cells; IHC-P: Human liver carcinoma tissue section.
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Liquid
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
pH: 7.40
Preservative: 0.02% Sodium azide
Constituents: 98.98% PBS, 1% BSA
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
Immunogen affinity purified
多克隆
IgG
Abpromise™承诺保证使用ab6046于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。
| 应用 | Ab评论 | 说明 |
|---|---|---|
| WB | (38) | 1/500. Detects a band of approximately 50 kDa (predicted molecular weight: 50 kDa). |
| ICC/IF | (11) | Use a concentration of 1 µg/ml. |
| IHC-P | (1) | Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
| IP | (1) | Use at an assay dependent concentration. |
Entrez Gene: 396254 Chicken
Entrez Gene: 203068 Human
Entrez Gene: 22154 Mouse
Entrez Gene: 380418 Xenopus laevis
Entrez Gene: 386701 Zebrafish
Omim: 191130 Human
SwissProt: P07437 Human
SwissProt: P99024 Mouse
SwissProt: Q91575 Xenopus laevis
Unigene: 636480 Human
Unigene: 46552 Xenopus laevis
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Immunocytochemistry/ Immunofluorescence - Anti-beta Tubulin antibody - Loading Control (ab6046)
ab6046 staining beta Tubulin in HeLa cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab6046 at 1µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).
Also suitable in cells fixed with 100% methanol (5 min).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
Western blot - Anti-beta Tubulin antibody - Loading Control (ab6046)
All lanes : Anti-beta Tubulin antibody - Loading Control (ab6046) at 1/500 dilution
Lane 1 : HeLa Cell lysate
Lane 2 : A431 Cell lysate
Lane 3 : MCF7 Cell lysate
Lane 4 : 293 Cell lysate
Lane 5 : HeLa Cell lysate with Human beta Tubulin peptide (ab20775) at 1 µg/ml
Lane 6 : A431 Cell lysate with Human beta Tubulin peptide (ab20775) at 1 µg/ml
Lane 7 : MCF7 Cell lysate with Human beta Tubulin peptide (ab20775) at 1 µg/ml
Lane 8 : 293 Cell lysate with Human beta Tubulin peptide (ab20775) at 1 µg/ml
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 50 kDa
Observed band size: 55 kDawhy is the actual band size different from the predicted?
Exposure time: 10 seconds
Western blot - Anti-beta Tubulin antibody - Loading Control (ab6046)
All lanes : Anti-beta Tubulin antibody - Loading Control (ab6046) at 1 µg/ml
Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : NIH3T3 (Mouse embryo fibroblast cell line) whole cell lysate
Lane 3 : PC12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
Lane 4 : CHO/K1 (Chinese hamster ovary cell line) whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/50000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 50 kDa
Observed band size: 51 kDawhy is the actual band size different from the predicted?
Exposure time: 30 seconds
Immunoprecipitation - Anti-beta Tubulin antibody - Loading Control (ab6046)
Beta Tubulin was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to Tubulin and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab6046.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 50kDa: beta Tubulin.
Immunocytochemistry/ Immunofluorescence - Anti-beta Tubulin antibody - Loading Control (ab6046)
ICC/IF image of ab6046 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then incubated in 1% BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab6046, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab150081 Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
The negative control (inset) is a secondary-only assay to demonstrate low non-specific binding of the secondary antibody.
This product also gave a positive signal under the same testing conditions in HeLa cells fixed with 100% methanol (5 min).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Tubulin antibody - Loading Control (ab6046)
IHC image of beta Tubulin staining in human liver carcinoma FFPE section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab6046, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
